Application of resonance raman microscopy to in vivo carotenoid.

Abstract

The high antioxidant activity of astaxanthin has been attracted considerable attention in these days. One of the major antioxidant activities of this carotenoid is anti-photoaging. We have been focusing our attention on this particular issue. The anti-photoaging activity should be functioning in inner skin. In this study we tried to find out the fact that astaxanthin that has been swabbed on the outer surface of the skin has really passed through and reached to the inner skin. For this purpose resonance Raman microscopy was applied to the rat skin sample on which astaxanthin was swabbed on its outer surface. Astaxanthin gives rise to a unique Raman spectrum that is characteristic of its molecular structure. Therefore, we can easily identify the presence or absence of astaxanthin in the area of the rat skin that is subjected to this spectroscopic measurement. We used 532 nm laser light for probing the resonance Raman scattering of astaxanthin. Astaxanthin shows three strong Raman lines at 1508, 1145, and 993 cm(-1). These three lines are ascribable to the C=C stretching, C-C stretching, and C-CH(3) in-plane rocking vibrational modes, respectively. We have constructed confocal Raman microscope that has the spatial resolution of ca. 500 nm. Three-dimensional mapping of the Raman spectrum of astaxanthin has been performed in order to determine its distribution in the rat skin.